The heart has a complex, 3D structure comprised of sheets of aligned, elongated cardiomyocytes which form a transmural helical structure. The 3D microenvironment of the heart has been shown to assist in the development of cardiomyocytes and the 3D helical structure of the heart is crucial to appropriate function. However, current tissue engineering strategies are unable to precisely control 3D tissue architecture in cell- dense tissues in vitro. Thus, we aim to engineer novel 3D cardiac tissue which can recapitulate the anisotropic, helical heart structure by utilizing nanofabrication-based cell sheet engineering and human pluripotent stem cell-derived cardiomyocytes (hPS-CMs). To do so, we will utilize a developed platform consisting of a thermoresponsive, nanofabricated substratum (TNFS) and a gel casting method to fabricate 3D, scaffold-free tissues with layer-by-layer structural control. We will first engineer and characterize multilayered cardiac tissues as an improved heart-in-a-dish model. We will then determine the microenvironmental effects on stem cell-derived cardiomyocyte maturity, such as biomimetic nanotopographical cues, as well as the 3D cell-dense cardiac microenvironment. Finally, the 3D stacked tissues will be fabricated with varying structures and analyzed for contractile and electrical function as physiological models of myocardial tissue. These models will be used to investigate the structure-function relationship of human cardiac tissue.